Design, synthesis, and computational studies of novel pyrazoline-based dual EGFR/HER-2 inhibitors with apoptotic antiproliferative activity

• A series of new pyrazoline-based derivatives was designed and synthesised. • The new compounds were evaluated as antiproliferative agent targeting EGFR and HER-2. • Antiproliferative activities were evaluated against four human cancer cell lines. • Molecular docking studies were carried out against EGFR and Her-2 enzymes. • In silico ADME/ pharmacokinetic analysis was considered. The simultaneous targeting of EGFR and HER2 constitutes a legitimate anticancer strategy for the treatment of solid tumors. In response, we developed and synthesized a new group of pyrazoline compounds that act as dual-target inhibitors for EGFR and HER-2. The structure of the newly synthesized compounds was validated with 1 H NMR, 13 C NMR, and elemental analysis. The novel compounds' antiproliferative efficacy was evaluated against four cancer cell lines. All compounds showed GI 50 values ranging from 23 to 66 nM, with the breast cancer cell line (MCF-7) showing the highest sensitivity. Compounds 7c, 7d, 7f, 7h, 7j , and 7l had the strongest antiproliferative activity, with derivatives 7d, 7h , and 7j outperforming erlotinib in all tested cancer cell lines. The study revealed that compounds 7d and 7h are the most effective dual-target inhibitors of EGFR and HER-2, exceeding the reference EGFR inhibitor erlotinib and having comparable action to the clinically used HER-2 medication Lapatinib. We tested the apoptotic potential of 7d and 7h and found that both compounds cause apoptosis by turning on caspase-3, caspase-8, and Bax and decreasing the expression of Bcl-2, a protein that prevents apoptosis. Molecular docking studies revealed robust interactions of these compounds within the EGFR and HER-2 binding pockets, supported by molecular dynamics simulations that confirmed their stability. ADME profiling highlighted the pharmacokinetic advantages of these compounds, particularly 7h , as orally bioavailable and effective inhibitors. These findings suggest that pyrazoline-based inhibitors could serve as a foundation for future development of dual-targeted therapies to overcome resistance in cancer treatment.